We will utilize polymeric phospholipid membranes to prepare a new generation of biochemical separation matrices that exhibit marked improvements in selectivity, stability, reproducibility and tenability. We will focus on two primary objectives: design and implementation of a) membrane protein functionalized stationary phase materials and b) ion channel functionalized detectors for microchip separations. To achieve these goals, silica particles (ranging in diameter from 0.5 to 5 <m) and/or the interior walls of silica capillaries will be coated with highly- stabilized, biologically-functionalized phospholipid bilayers (PLB) to facilitate highly selective identification of transmembrane protein modulators existing in complex mixtures. Ion channel functionalized PLBs will be prepared on a microfabricated aperture embedded in the flow channels of a microfabricated chip to identify ion channel modulators from complex solutions. The PLB serves as the matrix for chemically and biologically functionalizing the silica matrices via diverse phospholipid headgroups and/or incorporation of membrane-associated and membrane-spanning proteins and receptors. The high degree of physical and chemical stability of the PLB coatings, imparted via formation of a polymer scaffold, will significantly improve the long-term stability and thereby applicability of stabilized PLB stationary phases. Incorporation of membrane proteins into the stabilized PLB stationary phases will provide a novel basis for separation and identification of physiologically and pharmacologically important analytes. In addition to separations, functionalized PLBs can be used for biophysical analysis of novel ligands, e.g. peptide based pharmaceuticals to identify structurally important features of the membrane. Successful realization of the research goals presented herein will prove useful for qualitative and quantitative analysis of combinatorial libraries, identification and verification of pharmaceutical targets, elucidation of biochemical pathways and novel binding interactions, and many others. PUBLIC HEALTH RELEVANCE: This research project will develop and implement new, state-of-the-art technologies for biochemical separations, allowing for identification of novel pharmacological and physiological regulators of biological function. This research project will also provide key enabling technologies that lead to a more efficient elucidation of key biochemical and biophysical parameters for the design of next generation peptide-based drugs for treatment of a range of disease states.